Dual phase multiplex polymerase chain reaction
Abstract
Highly specific and sensitive methods were developed for multiplex amplification of nucleic acids on supports such as microarrays. Based on a specific primer design, methods include five types of amplification that proceed in a reaction chamber simultaneously. These relate to four types of multiplex amplification of a target DNA on a solid support, directed by forward and reverse complex primers immobilized to the support and a fifth type--pseudo-monoplex polymerase chain reaction (PCR) of multiple targets in solution, directed by a single pair of unbound universal primers. The addition of the universal primers in the reaction mixture increases the yield over the traditional "bridge" amplification on a solid support by approximately ten times. Methods that provide multitarget amplification and detection of as little as 0.45-4.5.times.10.sup.-12 g (equivalent to 10.sup.2-10.sup.3 genomes) of a bacterial genomic DNA are disclosed.
- Inventors:
-
- Charlottesville, VA
- Darien, IL
- Issue Date:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 985411
- Patent Number(s):
- 7432055
- Application Number:
- 10/794,381
- Assignee:
- UChicago Argonne LLC (Chicago, IL)
- Patent Classifications (CPCs):
-
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
- DOE Contract Number:
- W-31-109-ENG-38
- Resource Type:
- Patent
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Pemov, Alexander, and Bavykin, Sergei. Dual phase multiplex polymerase chain reaction. United States: N. p., 2008.
Web.
Pemov, Alexander, & Bavykin, Sergei. Dual phase multiplex polymerase chain reaction. United States.
Pemov, Alexander, and Bavykin, Sergei. Tue .
"Dual phase multiplex polymerase chain reaction". United States. https://www.osti.gov/servlets/purl/985411.
@article{osti_985411,
title = {Dual phase multiplex polymerase chain reaction},
author = {Pemov, Alexander and Bavykin, Sergei},
abstractNote = {Highly specific and sensitive methods were developed for multiplex amplification of nucleic acids on supports such as microarrays. Based on a specific primer design, methods include five types of amplification that proceed in a reaction chamber simultaneously. These relate to four types of multiplex amplification of a target DNA on a solid support, directed by forward and reverse complex primers immobilized to the support and a fifth type--pseudo-monoplex polymerase chain reaction (PCR) of multiple targets in solution, directed by a single pair of unbound universal primers. The addition of the universal primers in the reaction mixture increases the yield over the traditional "bridge" amplification on a solid support by approximately ten times. Methods that provide multitarget amplification and detection of as little as 0.45-4.5.times.10.sup.-12 g (equivalent to 10.sup.2-10.sup.3 genomes) of a bacterial genomic DNA are disclosed.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Oct 07 00:00:00 EDT 2008},
month = {Tue Oct 07 00:00:00 EDT 2008}
}
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